LOCUS       sequence-459089-        6507 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6507)
  AUTHORS   Yu H, Ye X, Guo N, Nathans J
  TITLE     Frizzled 2 and frizzled 7 function redundantly in convergent 
            extension and closure of the ventricular septum and palate: evidence
            for a network of interacting genes.
  JOURNAL   Development. 2012 Dec 1;139(23):4383-94. doi: 10.1242/dev.083352. 
            Epub 2012 Oct 24.
  PUBMED    23095888
REFERENCE   2  (bases 1 to 6507)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1242/dev.083352"; journalName: "Development"; date: "2012-12-1- 
            1"; volume: "139"; issue: "23"; pages: "4383-94"
COMMENT     Sequence Label: pRK5-mFzd5-1D4
FEATURES             Location/Qualifiers
     source          1..6507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    76..97
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(112..117,118..135,136..142)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    150..166
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     155..177
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     174..190
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     174..190
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        219..598
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        599..802
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     753..773
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     799..823
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1033..1051
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1033..1050
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1064..1083
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1139..1155
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1139..1155
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     regulatory      1165..1174
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2978..2994)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2979..2995)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     regulatory      3013..3022
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3026..3160
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3063..3082)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3117..3136
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3229..3586
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3229..3420
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3252..3272)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3437..3572
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3606..3623)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3606..3622)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3615..3637)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3835..4290
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3922..3941)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4132..4153
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4307..4326)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4425..4447
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4485..4503)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4571..4675
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4676..4744,4745..5536)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4894..4913)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5707..6295
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6196..6215
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6449..6466
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
       61 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
      121 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
      181 cagctatgac atgattacga attaattcga gctcgcccga cattgattat tgactagtta
      241 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      301 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      361 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      421 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      481 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      541 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      601 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      661 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      721 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      781 ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc
      841 acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgcg gccgggaacg
      901 gtgcattgga acgcggattc cccgtgccaa gagtgacgta agtaccgcct atagagtcta
      961 taggcccacc cccttggctt cgttagaacg cggctacaat taatacataa ccttatgtat
     1021 catacacata cgatttaggt gacactatag aataacatcc actttgcctt tctctccaca
     1081 ggtgtccact cccaggtcca actgcacctc ggttctatcc caccgcggtg gcggccgctc
     1141 tagaactagt ggatcaattc cgccgccacc atggctcgac ccgacccgtc tgcgcctccc
     1201 tctcttctgc tgctgttgct ggcgcagctg gtgggccggg cagcggccgc ctccaaggcc
     1261 ccggtgtgcc aggaaatcac ggtgcccatg tgccgaggca tcggctacaa cctgacgcac
     1321 atgcccaacc agttcaacca tgacacgcag gacgaagcag gcctggaggt gcaccaattc
     1381 tggccgcttg tggagatcca ctgctcaccg gacctgcgct tcttcctgtg ctctatgtac
     1441 acgcccatct gtttgcctga ctaccacaag ccgctaccac cgtgccgttc cgtgtgcgag
     1501 cgcgccaagg ccggctgctc gccgctcatg cgccagtacg gcttcgcctg gcccgagcgc
     1561 atgagctgcg accgcctccc tgtgctgggc ggcgacgccg aggttctgtg tatggattat
     1621 aaccgaagcg aagccaccac cgcgtcccct aagtccttcc cggccaaacc tacactccca
     1681 ggaccaccag gggcgccatc ttccgggggc gagtgcccct cgggaggccc atccgtgtgc
     1741 acgtgccgcg agcccttcgt gcccatcctg aaggagtcac acccactcta caacaaggtg
     1801 cgcaccggcc aagtgcccaa ctgcgcggtg ccctgctacc agccgtcctt cagcccggac
     1861 gagcgcacgt tcgccacctt ctggattggc ctgtggtctg tgctgtgctt catctccacg
     1921 tccaccaccg ttgccacctt cctcattgac atggaacgat tccgctaccc tgagcgcccc
     1981 atcatcttct tgtctgcgtg ctacctgtgt gtgtcactgg gattcttggt gcgcctggta
     2041 gtgggccatg ccagcgtcgc ttgcagccgt gagcacagcc acattcacta tgagactacc
     2101 ggccctgcgc tgtgcacggt tgtcttcctc ttagtctatt tctttggcat ggccagctcc
     2161 atctggtggg tcatcctgtc gctcacctgg ttcttggcgg ctggcatgaa gtggggcaat
     2221 gaagccatcg caggttatgc acagtacttc caccttgctg cctggctcat ccccagtgtc
     2281 aagtccatta cggcgctggc actgagctcg gtggacgggg acccagtggc tggcatctgc
     2341 tatgtgggca accaaaacct gaactcacta cgaggctttg tcttgggccc actggtgctg
     2401 tacctgttgg tgggcacgct cttccttctg gcaggcttcg tgtcactctt ccgcatccgg
     2461 agcgtcatca agcagggtgg cactaagacg gacaagctag agaagctcat gatccgcatc
     2521 ggcatcttca ccctgctcta cacggtgcca gccagcatcg tggtggcctg ctacctgtat
     2581 gagcagcact accgggagag ctgggaggca gccctcacct gcgcgtgtcc gggaccggac
     2641 gctggccagc cacgcgccaa acccgagtac tgggtgctca tgctcaagta cttcatgtgc
     2701 ctggtggtgg gcatcacgtc gggagtctgg atctggtccg gcaagactct ggagtcttgg
     2761 cggcggttca ccagccgctg ctgctgcagc tctcggcggg gccacaagag cggtggcgct
     2821 atggccgcag gagactatgc ggaggccagc gccgcgctca ccggcaggac cgggccgcct
     2881 ggccccaccg ccgcatacca caagcaagtg tccctgtcgc acgtaggcgc gccttccaag
     2941 acagagacca gccaagtggc gcctgcctaa agcttatcga taccgtcgac ctcgaggggg
     3001 ggcccgagct tggccgccat ggcccaactt gtttattgca gcttataatg gttacaaata
     3061 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3121 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc gggaattaat tcggcgcagc
     3181 accatggcct gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa
     3241 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3301 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3361 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3421 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3481 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3541 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tgttaacagc
     3601 ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     3661 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     3721 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt
     3781 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     3841 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3901 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3961 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4021 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     4081 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4141 tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga
     4201 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4261 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     4321 atgccgcata gttaagccag ccccgacccc gccaacaccc gctgacgcgc cctgacgggc
     4381 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     4441 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     4501 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     4561 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4621 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4681 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4741 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4801 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4861 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4921 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4981 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5041 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5101 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5161 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5221 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5281 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5341 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5401 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5461 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5521 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5581 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5641 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5701 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5761 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5821 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5881 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5941 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6001 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6061 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6121 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6181 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6241 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6301 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6361 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     6421 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     6481 cccaatacgc aaaccgcctc tccccgc
//
